Effect of CUX1 on the Proliferation of Hu Sheep Dermal Papilla Cells and on the Wnt/β-Catenin Signaling Pathway

Hui Zhou; Sainan Huang; Xiaoyang Lv; Shanhe Wang; Xiukai Cao; Zehu Yuan; Tesfaye Getachew; Joram M Mwacharo; Aynalem Haile; Kai Quan; Yutao Li; Antonio Reverter; Wei Sun

doi:10.3390/genes14020423

Effect of CUX1 on the Proliferation of Hu Sheep Dermal Papilla Cells and on the Wnt/β-Catenin Signaling Pathway

Genes (Basel). 2023 Feb 7;14(2):423. doi: 10.3390/genes14020423.

Authors

Hui Zhou^{1

2}, Sainan Huang^{1

2}, Xiaoyang Lv^{2

3}, Shanhe Wang^{1

2}, Xiukai Cao^{2

3}, Zehu Yuan^{2

3}, Tesfaye Getachew⁴, Joram M Mwacharo⁴, Aynalem Haile⁴, Kai Quan⁵, Yutao Li⁶, Antonio Reverter⁶, Wei Sun^{1

2

3

7}

Affiliations

¹ College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.
² International Joint Reserarch Laboratory in Universities of Jiangsu Province of China for Domestic Animal Germplasm Resources and Gentic Improvement, Yangzhou University, Yangzhou 225009, China.
³ Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou 225009, China.
⁴ International Centre for Agricultural Research in the Dry Areas, Addis Ababa 999047, Ethiopia.
⁵ College of Animal Science and Technology, Henan University of Animal Husbandry and Economics, Zhengzhou 450046, China.
⁶ CSIRO Agriculture and Food, 306 Carmody Rd, St Lucia, QLD 4067, Australia.
⁷ "Innovative China" "Belt and Road" International Agricultural Technology Innovation Institute for Evaluation, Protection, Improvement on Sheep Genetic Resource, Yangzhou 225009, China.

Abstract

CUT-like homeobox 1 protein (CUX1), also called CUX, CUTL1, and CDP, is a member of the DNA-binding protein homology family. Studies have shown that CUX1 is a transcription factor that plays an important role in the growth and development of hair follicles. The aim of this study was to investigate the effect of CUX1 on the proliferation of Hu sheep dermal papilla cells (DPCs) to reveal the role of CUX1 in hair follicle growth and development. First, the coding sequence (CDS) of CUX1 was amplified by PCR, and then CUX1 was overexpressed and knocked down in DPCs. A Cell Counting Kit-8 (CCK8), 5-ethynyl-2-deoxyuridine (EdU), and cell cycle assays were used to detect the changes in the proliferation and cell cycle of DPCs. Finally, the effects of overexpression and knockdown of CUX1 in DPCs on the expression of WNT10, MMP7, C-JUN, and other key genes in the Wnt/β-catenin signaling pathway were detected by RT-qPCR. The results showed that the 2034-bp CDS of CUX1 was successfully amplified. Overexpression of CUX1 enhanced the proliferative state of DPCs, significantly increased the number of S-phase cells, and decreased the number of G0/G1-phase cells (p < 0.05). CUX1 knockdown had the opposite effects. It was found that the expression of MMP7, CCND1 (both p < 0.05), PPARD, and FOSL1 (both p < 0.01) increased significantly after overexpression of CUX1 in DPCs, while the expression of CTNNB1 (p < 0.05), C-JUN, PPARD, CCND1, and FOSL1 (all p < 0.01) decreased significantly. In conclusion, CUX1 promotes proliferation of DPCs and affects the expression of key genes of the Wnt/β-catenin signaling pathway. The present study provides a theoretical basis to elucidate the mechanism underlying hair follicle development and lambskin curl pattern formation in Hu sheep.

Keywords: CUT-like homeobox 1 protein; Hu sheep; Wnt/β-catenin; dermal papilla cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Proliferation
Cells, Cultured
Hair Follicle
Matrix Metalloproteinase 7* / metabolism
Matrix Metalloproteinase 7* / pharmacology
Sheep
Wnt Signaling Pathway*

Substances

Matrix Metalloproteinase 7

Grants and funding

This work was supported by the National Natural Science Foundation of China-CGIAR (32061143036), National Natural Science Foundation of China (31872333, 32172689), Major New Varieties of Agricultural Projects in Jiangsu Province (PZCZ201739), High-end Foreign Expert Introduction Project (G2022014148L), Jiangsu 333 Distinguished Talents Project Foundation [(2022) 2-323], Natural Science Foundation of the Jiangsu Higher Education Institutions of China (22KJA230001), Major Project of Natural Science Foundation of Xinjiang Uyghur Autonomous Region [2022D01D47], Distinguished Talents Project Foundation of Yangzhou University, Jiangsu Natural Science Foundation for Colleges and Universities [21KJB230005], and Jiangsu Postdoctoral Science Foundation [2021K608C].